title: DNA microarray-based solid-phase RT-PCR for rapid detection and identification of influenza virus type A and subtypes H5 and H7.
abstract: Endemic of avian influenza virus (AIV) in Asia and epizootics in some European regions have caused considerable public concern on a possible pandemic of AIV. A rapid method for virus detection and effective surveillance in wild avian, poultry production as well as in humans is required. In this article, a DNA microarray-based solid-phase polymerase chain reaction (PCR) approach has been developed for rapid detection of influenza virus type A and for simultaneous identification of pathogenic virus subtypes H5 and H7. This solid-phase RT-PCR method combined reverse-transcription amplification of RNA extract in the liquid phase with sequence-specific nested PCR on the solid phase. A simple ultraviolet cross-linking method was used to immobilize the DNA probes over an unmodified glass surface, which makes solid-phase PCR a convenient possibility for AIV screening. The testing of 33 avian fecal and tracheal swab specimens was completed in less than 2 h with 94% accuracy.
authors.affiliation: DTU Nanotech, Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, DK-2800 Kgs. Lyngby, Denmark.
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
authors.forename: Raghuram
authors.initials: R
authors.lastname: Dhumpa
authors.forename: Dang Duong
authors.initials: DD
authors.lastname: Bang
authors.forename: Kurt
authors.initials: K
authors.lastname: Handberg
authors.forename: Anders
authors.initials: A
authors.lastname: Wolff
mesh_terms: D000818:Animals; D001717:Birds; D004587:Electrophoresis, Agar Gel; D005243:Feces / Q000821:virology; D009980:Influenza A virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000821:virology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014132:Trachea / Q000821:virology; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D012367:RNA, Viral; D014763:Viral Matrix Proteins